一步法快速检测食品中大肠菌群
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摘要
目的建立一步法快速检测食品中大肠菌群的斱法。方法食品参照GB 4789.3-2016进行前处理后,放入到大肠菌群检测试剂盒的样品制备瓶内,振摇制成样品匀液。将样品匀液1 mL加入到检测瓶中,配套使用在食源性致病菌同步检测仪中培养检测。结果在市场上购买3份样品用该斱法检测,幵同GB 4789.3-2016检测结果对比, 2种斱法吻合率高,且本斱法大肠菌群检测限能够达到10 CFU/g或mL, 70～480 min内可获得检测结果。结论该斱法可以实现对大肠菌群的快速检测,且操作简便,结果准确,为食品卫生质量监管提供参考。
Objective To establish a method for the rapid determination of coliform bacteria in foods by one-step method. Methods After pre-treating the food according to GB 4789.3-2016, the samples were put into preparation bottle of the coliform detection kit and were made to uniform liquid after shaking. One milliliter of the homogenate of the sample was added to the test bottle, and combined with the the synchronous detection instrument of foodborne pathogenic bacteria for detecting the coliform group. Results Three samples purchased in the market were detected by this method, and compared with the test results by GB 4789.3-2016, the anastomosis rate of the two methods was high, and the limit of detection of coliform group of this method could reach 10 CFU/g(mL), and the test results could be obtained within 70～480 minutes. Conclusion This method can realize the rapid detection of coliform group, the operation is simple and the result is accurate, which provides reference for the supervision of food hygiene quality.
Objective To establish a method for the rapid determination of coliform bacteria in foods by one-step method. Methods After pre-treating the food according to GB 4789.3-2016, the samples were put into preparation bottle of the coliform detection kit and were made to uniform liquid after shaking. One milliliter of the homogenate of the sample was added to the test bottle, and combined with the the synchronous detection instrument of foodborne pathogenic bacteria for detecting the coliform group. Results Three samples purchased in the market were detected by this method, and compared with the test results by GB 4789.3-2016, the anastomosis rate of the two methods was high, and the limit of detection of coliform group of this method could reach 10 CFU/g(mL), and the test results could be obtained within 70～480 minutes. Conclusion This method can realize the rapid detection of coliform group, the operation is simple and the result is accurate, which provides reference for the supervision of food hygiene quality.
引文
[1]潘慧华,李晶晶,刘坚真.大肠菌群一步发酵法检测的研究[J].食品科学,2006,27(12):637-641.Pan HH,Li JJ,Liu JZ.Studies on the detection of coliform bacteria by one-step fermentation[J].Food Sci,2006,27(12):637-641.
[2]杜连平,路福平.微生物学实验技术[M].北京:中国轻工业出版社,2008.Du LP,Lu FP.Microbiological experimental techniques[M].Beijing:China Light Industry Press,2008.
[3]卢勉飞,姚华卓,腾昆仑.蜡样芽孢杆菌显色培养基检测效果初步评价[J].食品科技,2013,38(6):313-317.Lu MF,Yao HZ,Teng KL.Preliminary evaluation on the detection effect of color medium of Bacillus cereus[J].Food Sci Technol,2013,38(6):313-317.
[4]刘慧.现代食品微生物学[M].北京:中国轻工业出版社,2006.Liu H.Modern food microbiology[M].Beijing:China Light Industry Press,2006.
[5]陈映灿.大肠菌群快速检测培养基的研制及评价[D].郑州:郑州大学,2012.Chen YC.Development and evaluation of rapid detection medium for coliform bacteria[D].Zhengzhou:Zhengzhou University,2012.
[6]GB 4789.38-2012食品微生物学检验大肠埃希氏菌计数[S].GB 4789.38-2012 Food microbiological analysis-Escherichia coli count[S].
[7]Geoffrey N,Jonathan N.Method and apparatus utilizing enzyme substrates producing slow diffusing fluorescent product appearances and chromogens,and use in combination with fast diffusing product appearances:US,20160017402[P].2016-01-21.
[8]Hofmann J,Sernetz M.Immobilized enzyme kinetics analyzed by flow-through microfluorimetry:Resorufin-β-D-galactopyranoside as a new fluorogenic substrate forβ-galactosidase[J].Anal Chim Acta,1984,163:67-72.
[9]GB 4789.3-2016食品微生物学检验大肠菌群计数[S].GB 4789.3-2016 Food microbiological analysis-Enumeration of coliforms[S].
[10]GB 4789.3-2010食品微生物学检验大肠菌群计数[S].GB 4789.3-2010 Food microbiological analysis-Enumeration of coliforms[S].
[11]马延霞,吴清平,张菊梅.大肠菌群特异性检测荧光底物MUGal的合成及应用[J].现代食品科技,2014,30(8):251-257.Ma YX,Wu QP,Zhang JM.Synthesis and application of coliform specific detection of fluorescent substrate MUGal[J].Mod Food Sci Technol,2014,30(8):251-257.
[12]林露,严维凌,朱祖琪,等.实时荧光PCR技术快速检测肉制品中肉含量[J].中国食品学报,2016,16(3):152-159.Lin Lu,Yan WL,Zhu ZQ,et al.Rapid determination of meat content in meat products by real-time fluorescence PCR[J].J Chin Inst Food Sci Technol,2016,16(3):152-159.
[13]James DM,Hendrson F.Nucleoside triphosphate specifi of firefly luciferase[J].Anal Biochemm,1983,131(1):187-189.
[14]李红.显色培养基法快速检测大肠菌群和大肠杆菌的效果分析[J].中国冶金工业医学杂志,2017,34(6):665.Li H.Effect analysis of rapid detection of coliform and escherichia coli by chromogenic medium method[J].Chin Med J Metal Ind,2017,34(6):665.
[15]吴清平,马延霞,张菊梅,等.香豆素类荧光底物的合成及其在微生物检测中的应用进展[J].化工进展,2014,33(9):2444-2449.Wu QP,Ma YX,Zhang JM,et al.Synthesis of coumarin fluorescent substrates and their application in microbiological detection[J].Chem Prog,2014,33(9):2444-2449.
[2]杜连平,路福平.微生物学实验技术[M].北京:中国轻工业出版社,2008.Du LP,Lu FP.Microbiological experimental techniques[M].Beijing:China Light Industry Press,2008.
[3]卢勉飞,姚华卓,腾昆仑.蜡样芽孢杆菌显色培养基检测效果初步评价[J].食品科技,2013,38(6):313-317.Lu MF,Yao HZ,Teng KL.Preliminary evaluation on the detection effect of color medium of Bacillus cereus[J].Food Sci Technol,2013,38(6):313-317.
[4]刘慧.现代食品微生物学[M].北京:中国轻工业出版社,2006.Liu H.Modern food microbiology[M].Beijing:China Light Industry Press,2006.
[5]陈映灿.大肠菌群快速检测培养基的研制及评价[D].郑州:郑州大学,2012.Chen YC.Development and evaluation of rapid detection medium for coliform bacteria[D].Zhengzhou:Zhengzhou University,2012.
[6]GB 4789.38-2012食品微生物学检验大肠埃希氏菌计数[S].GB 4789.38-2012 Food microbiological analysis-Escherichia coli count[S].
[7]Geoffrey N,Jonathan N.Method and apparatus utilizing enzyme substrates producing slow diffusing fluorescent product appearances and chromogens,and use in combination with fast diffusing product appearances:US,20160017402[P].2016-01-21.
[8]Hofmann J,Sernetz M.Immobilized enzyme kinetics analyzed by flow-through microfluorimetry:Resorufin-β-D-galactopyranoside as a new fluorogenic substrate forβ-galactosidase[J].Anal Chim Acta,1984,163:67-72.
[9]GB 4789.3-2016食品微生物学检验大肠菌群计数[S].GB 4789.3-2016 Food microbiological analysis-Enumeration of coliforms[S].
[10]GB 4789.3-2010食品微生物学检验大肠菌群计数[S].GB 4789.3-2010 Food microbiological analysis-Enumeration of coliforms[S].
[11]马延霞,吴清平,张菊梅.大肠菌群特异性检测荧光底物MUGal的合成及应用[J].现代食品科技,2014,30(8):251-257.Ma YX,Wu QP,Zhang JM.Synthesis and application of coliform specific detection of fluorescent substrate MUGal[J].Mod Food Sci Technol,2014,30(8):251-257.
[12]林露,严维凌,朱祖琪,等.实时荧光PCR技术快速检测肉制品中肉含量[J].中国食品学报,2016,16(3):152-159.Lin Lu,Yan WL,Zhu ZQ,et al.Rapid determination of meat content in meat products by real-time fluorescence PCR[J].J Chin Inst Food Sci Technol,2016,16(3):152-159.
[13]James DM,Hendrson F.Nucleoside triphosphate specifi of firefly luciferase[J].Anal Biochemm,1983,131(1):187-189.
[14]李红.显色培养基法快速检测大肠菌群和大肠杆菌的效果分析[J].中国冶金工业医学杂志,2017,34(6):665.Li H.Effect analysis of rapid detection of coliform and escherichia coli by chromogenic medium method[J].Chin Med J Metal Ind,2017,34(6):665.
[15]吴清平,马延霞,张菊梅,等.香豆素类荧光底物的合成及其在微生物检测中的应用进展[J].化工进展,2014,33(9):2444-2449.Wu QP,Ma YX,Zhang JM,et al.Synthesis of coumarin fluorescent substrates and their application in microbiological detection[J].Chem Prog,2014,33(9):2444-2449.